Abstract
Bone marrow stromal cells (BMSCs) for osteoblast differentiation studies can be obtained by gradient isolation techniques or by directly plating a filtered cell suspension. We compared these two procedures to evaluate whether this step is critical in order to obtain a high number of differentiated colonies. Isolated primary rat BMSCs were cultured in vitro with or without insulin-like growth factor II (IGFII), basic fibroblast growth factor (b-FGF), epidermal growth factor (EGF) or transforming growth factor β1 (TGFβ1), and histochemically and biochemically analysed at different time points. The gradient procedure produced a significantly higher number of colonies capable of osteoblastic differentiation. The growth factors had different effects. In particular, b-FGF and EGF significantly increased the number of Alizarin red S positive colonies, while IGFII and TGFβI exerted inhibitory effects. Nodules obtained on day 21 showed some alkaline phosphatase positive cells and were Von Kossa-positive. These data demonstrate that more differentiated colonies are obtainable from BMSCs isolated by the gradient procedure.
Original language | English |
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Pages (from-to) | 49-58 |
Number of pages | 10 |
Journal | Connective Tissue Research |
Volume | 42 |
Issue number | 1 |
Publication status | Published - 2001 |
Keywords
- Bone marrow stromal cells
- Differentiation
- Growth factors
- Isolation procedure
- Mineralization
ASJC Scopus subject areas
- Biochemistry
- Immunology
- Nephrology
- Cell Biology
- Orthopedics and Sports Medicine